Re: PI toxicity

Dennis_Young@CIS.ucsd.edu
Thu, 30 Nov 1995 19:44:00 -0800

I use PI at 0.5 micrograms per ml (in "plain" PBS)for live/dead
discrimination and at 50 mikes/ml with fixed cells for DNA cell cycle.


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* Dennis J. Young Voice : (619) 822-0407 *
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______________________________ Reply Separator _________________________________
Subject: PI toxicity
Author: DrBlood@medschool.med.utah.edu at @UCSD
Date: 11/29/95 9:40 AM

Judy the Gambler asked:

>> 1) Is PI toxic to cells?

*Everything* (including pizza) is toxic at some doses 8->. However, I
routinely use PI for live/dead discrimination on cell populations that are
subsequently used in functional assays. A PI concentration of 10 micrograms
per ml or so should be fine for maintaining viability of live cells. As I
understand it, the 7-AAD dye can not be used for viability staining without
harming the live cells, but PI is OK.

>> 2) What is the minimum incubation time for PI and cells, say for
live/dead discrimination or DNA?

It stains pretty fast, say 5 minutes or so should be saturating. We
usually just resuspend the cell population in PI and analyze directly. The
free PI has little fluorescence so it does not need to be washed out.

Jerry Spangrude



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